Gene and drug therapies are being developed to al eviate vision loss in patients with Stargardt’s disease and age-related macular degeneration （AMD）. To evaluate the therapeutic effects of these treatments, organic...Gene and drug therapies are being developed to al eviate vision loss in patients with Stargardt’s disease and age-related macular degeneration （AMD）. To evaluate the therapeutic effects of these treatments, organic sol-vents are routinely used to extract and quantify bisretinoid lipofuscin constituents, such as N-retinylidene-N-retinyl-ethanolamine （A2E） and all-trans-retinal dimer （ATR-dimer）. By high-performance liquid chromatography （HPLC）, we found that A2E and ATR-dimer were both altered by tetrahydrofuran （THF） and chloroform, but were stable in dimethyl sulfoxide （DMSO） or methanol （MeOH）. In addition, cyclohexane and ethanol （EtOH） did not alter ATR-dimer, whereas an alteration of A2E occurred in EtOH. On the basis of these findings, we designed processes II-IV, generated by modifications of process I, a routine method to measure bisretinoid compounds in vivo. Extra amounts of either ATR-dimer or A2E in mouse eyecups were released by processes II-IV versus process I. Efforts to clarify the effects of organic solvents on lipofuscin pigments are important because such studies can guide the handling of these fluoro-phores in related experiments.展开更多
基金Project supported by the National Natural Science Foundation of China(Nos.21202146 and 81271018)the Fundamental Research Funds for the Central Universities+1 种基金the Zhejiang Scientific Research Foundation for the Returned Overseas Chinese Scholars(No.J20120556)the Zhejiang Key Laboratory Fund of China(No.2011E10006)
文摘Gene and drug therapies are being developed to al eviate vision loss in patients with Stargardt’s disease and age-related macular degeneration （AMD）. To evaluate the therapeutic effects of these treatments, organic sol-vents are routinely used to extract and quantify bisretinoid lipofuscin constituents, such as N-retinylidene-N-retinyl-ethanolamine （A2E） and all-trans-retinal dimer （ATR-dimer）. By high-performance liquid chromatography （HPLC）, we found that A2E and ATR-dimer were both altered by tetrahydrofuran （THF） and chloroform, but were stable in dimethyl sulfoxide （DMSO） or methanol （MeOH）. In addition, cyclohexane and ethanol （EtOH） did not alter ATR-dimer, whereas an alteration of A2E occurred in EtOH. On the basis of these findings, we designed processes II-IV, generated by modifications of process I, a routine method to measure bisretinoid compounds in vivo. Extra amounts of either ATR-dimer or A2E in mouse eyecups were released by processes II-IV versus process I. Efforts to clarify the effects of organic solvents on lipofuscin pigments are important because such studies can guide the handling of these fluoro-phores in related experiments.
